Preparative reversed-phase high-performance liquid chromatography collection efficiency for an antimicrobial peptide on columns of varying diameters (1mm to 9.4mm I.D.)

Journal of Chromatography. a
Yuxin ChenR S Hodges

Abstract

The present study examines the effect of reversed-phase high-performance liquid chromatography (RP-HPLC) column diameter (1mm to 9.4mm I.D.) on the one-step slow gradient preparative purification of a 26-residue synthetic antimicrobial peptide. When taken together, the semi-preparative column (9.4mm I.D.) provided the highest yields of purified product (an average of 90.7% recovery from hydrophilic and hydrophobic impurities) over a wide range of sample load (0.75-200mg). Columns with smaller diameters, such as narrowbore columns (150x2.1mm I.D.) and microbore columns (150x1.0mm I.D.), can be employed to purify peptides with reasonable recovery of purified product but the range of the crude peptide that can be applied to the column is limited. In addition, the smaller diameter columns require more extensive fraction analysis to locate the fractions of pure product than the larger diameter column with the same load. Our results show the excellent potential of the one-step slow gradient preparative protocol as a universal method for purification of synthetic peptides.

References

Jun 15, 1992·Analytical Chemistry·J G DorseyH G Barth
Jul 12, 1991·Journal of Chromatography·R S HodgesC T Mant
Jan 1, 1983·Methods in Enzymology·F E Regnier
Jan 1, 1995·Biophysical Journal·S E BlondelleE Pérez-Payá
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Jan 29, 2005·The Journal of Biological Chemistry·Yuxin ChenRobert S Hodges
Feb 24, 2006·Chemical Biology & Drug Design·Yuxin ChenRobert S Hodges

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Citations

Dec 13, 2006·Antimicrobial Agents and Chemotherapy·Yuxin ChenRobert S Hodges
Apr 13, 2017·Journal of Chromatography. a·Carmen R M BriaS Kim Ratanathanawongs Williams
Dec 19, 2008·Chemical Biology & Drug Design·Ziqing JiangRobert S Hodges
Jul 27, 2012·Journal of Peptide Science : an Official Publication of the European Peptide Society·Paul W R HarrisMargaret A Brimble
Nov 26, 2014·Journal of Peptide Science : an Official Publication of the European Peptide Society·Tonghui YiYuxin Chen
Mar 20, 2014·Chemistry : a European Journal·Paul W R HarrisMargaret A Brimble
Jul 31, 2007·Journal of Chromatography. a·João Ezequiel OliveiraMaria Teresa Carvalho Pinto Ribela
Feb 11, 2010·Journal of Structural Biology·J Paul Kirwan, Robert S Hodges

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